草鱼自噬相关基因Beclin1的克隆及其在MC-LR胁迫下的表达特征

为评估微囊藻毒素-LR (MC-LR)对鱼类自噬的影响, 根据转录组测序结果得到的EST序列, 采用RACE技术获得了草鱼(Ctenopharygodon idella)自噬基因Beclin1 (CiBeclin1)的cDNA全长序列。该基因序列全长1590 bp, 包括1344 bp的开放阅读框, 编码447个氨基酸, 分子量为51.2 kD, 理论等电点为4.88。CiBeclin1包含1个BH3和1个ECD结构域。CiBeclin1氨基酸序列与其他物种的相似性为88%—98%, 构建的进化树显示CiBeclin1与稀有鲫(Gobiocypris rarus)的Beclin1亲缘关系最近。实时荧光定量PCR (qRT-PCR)检测结果表明, CiBeclin1在肝、肾、脾等10种不同组织中均广泛表达, 但在肝脏组织中的相对表达量最丰富, 显著高于相对表达量最低的头肾组织(P<0.05)。在不同剂量(25和100 μg MC-LR/kg BW) MC-LR胁迫24h、48h、72h和96h后, 草鱼肝脏中CiBeclin1表达量均显著低于对照组的表达水平, 研究结果表明, MC-LR能抑制草鱼CiBeclin1的表达, 但MC-LR是否在该剂量下诱导了草鱼的自噬还有待于进一步研究。     

全球中华鲎资源保护现状及对策建议

鲎是古老的海洋节肢动物。中华鲎(Tachypleus tridentatus)是世界现存4种鲎中体型最大的一种, 是河口生态系统的标志物种, 同时其血液被用于生产医用检验试剂――鲎试剂。中华鲎的自然地理分布范围相当狭窄, 仅局限于日本濑户内海向南延伸至印度尼西亚爪哇岛北岸以北的太平洋西岸海域, 其中在中国东岸和日本南部海域的历史产量较高。自20世纪50年代以来中华鲎种群数量出现了显著减少, 2019年中华鲎在IUCN红色名录中的濒危等级正式更新为濒危(EN), 明确了中华鲎资源呈现全球性衰退的状态, 究其原因可归纳为鲎生境破坏和过度捕捞两个方面。在开展鲎资源保护的实践工作中, 作者深刻反思当前鲎资源保护在海洋保护区划定、增殖放流及科普和野生动物保护法宣传中存在的问题并提出相应建议, 包括加快完善种群基线数据, 制定标准化种群和生境基线监测指南, 构建科学放流体系等, 以期推进全球范围内的中华鲎资源保护与科学管理。     

Estimation of Cellobiohydrolase I Activity by Numerical Differentiation of Dynamic Ultraviolet Spectroscopy

1,4-β-D-glucan cellobiohydrolase I (CBH I),p-nitrophenyl β-D-cellobioside,p-nitrophenol andcellobiose show distinct ultraviolet spectra,allowing the design of an assay to track the dynamic process ofp-nitrophenyl β-D-cellobioside hydrolysis by CBH I.Based on the linear relationship between p-nitrophenolformation in the hydrolysate and its first derivative absorption Curve of AUC340_400_(nm)(area under the curve),a new sensitive assay for the determination of CBH I activity was developed.The dynamic parameters ofcatalysis reaction,such as Vm and k_(cat),can all be derived from this result.The influence of β-glucosidase andendoglucanase in crude enzyme sample on the assay was discussed in detail.This approach is useful foraccurate determination of the activity of CBHs.     

水稻主茎节位分蘖及生产力补偿能力

本研究选用两个籼稻杂交品种西农优1号和冈优725,采用人工去蘖形成分蘖初期的早发、中发、迟发三个处理,在群体条件下研究分蘖和生产力的补偿能力。本研究发现,迟发处理高节位群的分蘖补偿能力最明显,但绝对补偿量小。迟发处理高节位群的生长势、有效分蘖倍数增加,抽穗期单茎叶面积有增加的趋势,但抽穗期有效穗数、叶面积系数降低;迟发处理冠层光分布增加,叶绿素、蛋白质含量提高,叶片细胞内保护酶类(Superoxide dismutase, SOD、catalase,CAT)活性下降慢,叶片衰老慢,但是漏光率增加,截光率下降;迟发处理净光合生产率（Net assimulation rate, NAR）增加,同化物分配合理,经济系数有增加的趋势,但物质积累速度（Crop growth rate ,CGR）和干物质生产量减少。在本实验条件下,早发与中发处理的经济产量差异不显著,但早发与迟发处理的经济产量差异达极显著。本研究结果表明大苗秧对生物、非生物因素胁迫,如本田前期一定程度的人工去蘖,具有一定的分蘖补偿能力。     

Plant biodiversity and its conservation strategy in the in-undation and resettlement districts of the Yangtze Three Gorges, China

The investigation on and conservation project of plant biodiversity in the inundation and resettlement districts of the Three Gorges region were carried out with the ongoing construction of the Three Gorges Dam. Based on the first-hand specimens collected for many years and the historical records, we found that there were about 1784 higher plant species, belonging to 762 genera of 170 families, which accounted for 59.19%, 75.30% and 85.85% of the total amount of species, genera and families in the Three Gorges region, including 27 species specific in these two districts. The shrub and grass communities were mostly distributed along the low-altitude river bank zone of the Yangtze River and were largely influenced by the dynamic water level of the Three Gorges reservoir area. The measurement of Shannon-Wiener and Pielou evenness indexes on 21 Pinus massoniana (Masson pine) communities distributed in the Three Gorges reservoir area was carried out, and results showed that the biodiversity of shrub layers was the highest, followed by grass layers, the middle, and tree layers, the lowest. Among the 11 typical shrub communities in this region, the abundance of shrub layers of Form. Rhus chinensis, Form. Cotinus coggygria var. cinerea, Form. Vitex negundo var. cannabifoliaand and Form. Coriaria sinica was relatively higher, which were 16, 26, 29 and 15, respectively. These four shrubs also showed high biodiversity index (H′), which were 1.791, 3.427, 2.949 and 1.718, respectively. Among the 9 grasslands mainly distributed along the river bank zone, 4 grassland communities with higher altitude (Form. Dicranopteris dichotoma, Form. Miscanthus floridutus, Form. Imperata cylindrica var. major and Form. Arthraxon hispidus) showed higher abundance and biodiversity, which were 1.697, 1.354, 1.144 and 1.018, respectively. Considering the inundation and resettlement districts, there were 22 vegetation types threatened by submersion, including 4 woody plant communities, 9 shrub communities and 9 grassland communities. The 6 shrub communities of Form. Securinega wuxiensis, Form. Bauhinia brachycarpa, Form. Geum aleppicum, Form. Distylium chinense, Form. Buxus henryi and Form. Myrsine africana would be fully submerged and part of Adianthum reniforme var. sinensis and Buxus ichangensis would be also submerged. At present, plant species conservation and monitoring stations in the Three Gorges region have been constructed, and more than 200 plant species have already been transplanted and conserved, including 37 threatened species listed in the China Plant Red Data Book and 11 local constructive species.     

Drought-resistant physiological characteristics of four shrub species in arid valley of Minjiang River, China

The formation of 161 km-long arid valley (170 km²) of Mingjiang River is attributed to the effect of foehn. The arid valley presents harsh conditions for the growth of vegetation, which include higher annual evaporation and transpiration rate (1400–2000 mm), lower annual precipitation rate (400–700 mm), steep slope, and infertile soils. Although the vegetation is dominated by xerophilous bushes and grasses, it plays a significant role in soil and water conservation in watershed and provides important references in the arid valley as it is very difficult to grow trees. Drought-resistant physiological characteristics of four native species, i.e., Sophora davidiana, Bauhinia faberi var. microphylla, Convolvulus tragacanthoides, and Artemisia gmelinii, have been measured. It was found that the content of chlorophyll, free proline and soluble sugars showed an increase, whereas evaporation ratio, WSD, RWC and cytolemma osmosis showed a decrease under drought stress. Integrated assessment of the four species indicated that the integrated drought resistance index was the highest in July (0.507), followed by September, August and June, whereas it was the lowest in May (0.442). The result provided a basis for vegetation restoration, especially it improved survival of planted species in May and June when plant species exhibit the weakest drought resistance.     

槐耳清膏抑制非小细胞肺癌细胞增殖和血管新生的作用机制研究

目的:探讨槐耳清膏抑制非小细胞肺癌(NSCLC)细胞增殖和血管新生的作用机制。方法:选择对数生长期的NSCLC细胞,经过传代培养成细胞株后采用随机法分成低剂量组、高剂量组以及空白对照组。其中低剂量组和高剂量组分别加入5μmol/L、10μmol/L的槐耳清膏进行处理,而空白对照组未加入槐耳清膏处理。利用细胞增殖毒性试验法(MTT)检测NSCLC细胞的存活率,并采用蛋白免疫印迹实验(WB)法检测表皮生长因子受体(EGFR)和血管内皮生长因子受体2(VEGFR2)、磷酸化EGFR(pEGFR)、磷酸化VEGFR2(pVEGFR2)、磷脂酰肌醇3-激酶(PI3K)、磷酸化丝氨酸/苏氨酸特异性蛋白激酶(pAKT)(pAKT)、磷酸化细胞外信号调节激酶1/2(pERK1/2)、磷酸化应激活化蛋白激酶1/2(pJNK1/2)、磷酸化-p38(pp38)、细胞周期蛋白D1(Cyclin D1)及增殖细胞核抗原(PCNA)蛋白表达水平。结果:低剂量组、高剂量组NSCLC细胞的存活率均显著低于空白对照组(P<0.05)。与空白对照组相比,槐耳清膏处理NSCLC细胞后,低剂量组、高剂量组中pEGFR、pVEGFR2蛋白的相对表达水平均显著降低(P<0.05)。与空白对照组比较,槐耳清膏处理NSCLC细胞后,低剂量组、高剂量组中PI3K蛋白、pAKT、pERK1/2、pJNK1/2、pp38、Cyclin D1及PCNA的相对表达水平均显著降低(P<0.05)。结论:基于EGFR/VEGFR2信号通路,槐耳清膏对NSCLC细胞增殖和血管新生有一定的抑制作用,可能成为一种针对NSCLC的有用靶向药物。     

iASPP protects the heart from ischemia injury by inhibiting p53 expression and cardiomyocyte apoptosis

Currently, there remains a great need to elucidate the molecular mechanism of acute myocardial infarction in order to facilitate the development of novel therapy. Inhibitor of apoptosis-stimulating protein of p53 (iASPP) is a member of the ASPP family proteins and an evolutionarily preserved inhibitor of p53 that is involved in many cellular processes, including apoptosis of cancer cells. The purpose of this study was to investigate the possible role of iASPP in acute myocardial infarction. The protein level of iASPP was markedly reduced in the ischemic hearts in vivo and hydrogen peroxide-exposed cardiomyocytes in vitro. Overexpression of iASPP reduced the infarct size and cardiomyocyte apoptosis of mice subjected to 24 h of coronary artery ligation. Echocardiography showed that cardiac function was improved as indicated by the increase in ejection fraction and fractional shortening. In contrast, knockdown of iASPP exacerbated cardiac injury as manifested by impaired cardiac function, increased infarct size, and apoptosis rate. Mechanistically, overexpression of iASPP inhibited, while knockdown of iASPP increased the expressions of p53 and Bax, the key regulators of apoptosis. Taken together, our results suggested that iASPP is an important regulator of cardiomyocyte apoptosis, which represents a potential target in the therapy of myocardial infarction.     

The Role of DNA Methylation Reprogramming During Sex Determination and Transition in Zebrafish

DNA methylation is a prevalent epigenetic modification in vertebrates, and it has been shown to be involved the regulation of gene expression and embryo development. However, it remains unclear how DNA methylation regulates sexual development, especially in species without sex chromosomes. To determine this, we utilized zebrafish to investigate DNA methylation reprogramming during juvenile germ cell development and adult female-to-male sex transition.We reveal that primordial germ cells(PGCs) undergo significant DNA methylation reprogramming during germ cell development, and the methylome of PGCs is reset to an oocyte/ovary-like pattern at 9 days post fertilization(9 dpf). When DNA methyltransferase(DNMT) activity in juveniles was blocked after 9 dpf, the zebrafish developed into females. We also show that Tet3 is involved in PGC development. Notably, we find that DNA methylome reprogramming during adult zebrafish sex transition is similar to the reprogramming during the sex differentiation from 9 dpf PGCs to sperm. Furthermore, inhibiting DNMT activity can prevent the female-to-male sex transition, suggesting that methylation reprogramming is required for zebrafish sex transition. In summary, DNA methylation plays important roles in zebrafish germ cell development and sexual plasticity.     

Call Characteristic Network Reveal Geographical Patterns of Call Similarity:Applying Network Analysis to Frog’s Call Research

Individual’s phenotypic traits are the results of adaptation to ecological conditions.Therefore,different selection pressures caused by heterogeneous environments may result in phenotypic difference,especially for individuals in different geographical populations.Here,we illustrated for the first time to use social network analysis(SNA)for examining whether geographical proximity predicts the similarity patterns in call characteristics among populations of an anuran species.We recorded calls from 150 male dorsal-striped opposite-fingered treefrogs(Chiromantis doriae)at 11 populations in Hainan Province and one population in Guangdong Province in China's Mainland,and we measured eight acoustic variables for each male.Mantel test didn’t show a correlation between geographical proximity and the similarity in call characteristics among populations.In addition,we failed to find correlations between a population’s eigenvector centrality and the distance to its nearest neighbor,nor between the coefficient of variation of similarity in call characteristics of a population and the average distance to all other populations.Nevertheless,three acoustic clusters were identified by the Girvan-Newman algorithm,and clustering was partially associated with geography.Furthermore,the most central populations were included in the same cluster,but the top betweenness populations were located within different clusters,suggesting that centrality populations are not necessary bridging between clusters.These results demonstrate the potential usefulness of the SNA toolbox and indicate that SNA helps to uncover the patterns that often overlooked in other analytical methods.By using SNA in frog’s call studies,researchers could further uncover the potential relationship in call characteristics between geographical populations,further reveal the effects of ecological factors on call characteristics,and probably enhance our understanding of the adaptive evolution of acoustic signals.